Recombinant DNA technology and genetic engineering has allowed for the introduction of foreign DNA sequences into cells for the expression of proteins of interest. However, obtaining high expression remains a challenge. Expression of important transgenes in a cell at high levels requires the ability to control the regulatory mechanisms governing expression. This requires suitable regulatory sequences that can function with the desired transgenes.
Production of recombinant proteins has been one of the major challenges within the biotechnological industry. Bacterial cells, in particular Escherichia coli, have been widely used as host cells for the production of recombinant polypeptides.
Polynucleotide compositions that provide enhanced gene expression provide certain benefits. These benefits include not only improved efficiency, cost-effectiveness, consistency and accuracy in improving the expression of certain genes, but also the ability to achieve a far greater scope of applicability. Current methods of enhancing gene expression include mutating regulator regions to increase transcription. In addition, mutation within the coding sequence can enhance protein stability. However, it would be desirable to have an approach to attain enhanced gene expression without having to alter a substantial number of codons of the gene or identify inhibitory sequences of the gene and then altering those sequences. The present invention provides a solution for the need in the art for enhancing gene expression.